CN109283025A - The detection method of contained polypeptide classes in a kind of pearl powder - Google Patents
The detection method of contained polypeptide classes in a kind of pearl powder Download PDFInfo
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- CN109283025A CN109283025A CN201811086157.5A CN201811086157A CN109283025A CN 109283025 A CN109283025 A CN 109283025A CN 201811086157 A CN201811086157 A CN 201811086157A CN 109283025 A CN109283025 A CN 109283025A
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- pearl powder
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- nacl solution
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- 239000000843 powder Substances 0.000 title claims abstract description 32
- 238000001514 detection method Methods 0.000 title claims abstract description 29
- 229920001184 polypeptide Polymers 0.000 title claims abstract description 28
- 102000004196 processed proteins & peptides Human genes 0.000 title claims abstract description 28
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 28
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims abstract description 34
- 239000003480 eluent Substances 0.000 claims abstract description 19
- 239000011780 sodium chloride Substances 0.000 claims abstract description 17
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 17
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims abstract description 13
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims abstract description 13
- 235000011130 ammonium sulphate Nutrition 0.000 claims abstract description 13
- 239000002244 precipitate Substances 0.000 claims abstract description 11
- 238000005185 salting out Methods 0.000 claims abstract description 11
- 239000007787 solid Substances 0.000 claims abstract description 10
- FEMOMIGRRWSMCU-UHFFFAOYSA-N ninhydrin Chemical compound C1=CC=C2C(=O)C(O)(O)C(=O)C2=C1 FEMOMIGRRWSMCU-UHFFFAOYSA-N 0.000 claims abstract description 8
- 238000000926 separation method Methods 0.000 claims abstract description 7
- CUXSAAMWQXNZQW-UHFFFAOYSA-N acetic acid;butan-1-ol Chemical compound CC(O)=O.CCCCO CUXSAAMWQXNZQW-UHFFFAOYSA-N 0.000 claims abstract description 5
- 239000002904 solvent Substances 0.000 claims abstract description 5
- 238000009835 boiling Methods 0.000 claims abstract description 4
- 238000001962 electrophoresis Methods 0.000 claims description 4
- 238000002791 soaking Methods 0.000 claims description 3
- 230000003712 anti-aging effect Effects 0.000 abstract description 6
- 230000000144 pharmacologic effect Effects 0.000 abstract description 4
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 230000003796 beauty Effects 0.000 description 2
- 230000007062 hydrolysis Effects 0.000 description 2
- 238000006460 hydrolysis reaction Methods 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 230000007812 deficiency Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 150000002469 indenes Chemical class 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000001114 myogenic effect Effects 0.000 description 1
- 230000002936 tranquilizing effect Effects 0.000 description 1
Classifications
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/38—Diluting, dispersing or mixing samples
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N1/00—Sampling; Preparing specimens for investigation
- G01N1/28—Preparing specimens for investigation including physical details of (bio-)chemical methods covered elsewhere, e.g. G01N33/50, C12Q
- G01N1/44—Sample treatment involving radiation, e.g. heat
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
- G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
- G01N21/31—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry
- G01N21/33—Investigating relative effect of material at wavelengths characteristic of specific elements or molecules, e.g. atomic absorption spectrometry using ultraviolet light
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
- G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
- G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
- G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
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- Physics & Mathematics (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Physics & Mathematics (AREA)
- Pathology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Analytical Chemistry (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Spectroscopy & Molecular Physics (AREA)
- Engineering & Computer Science (AREA)
- Plasma & Fusion (AREA)
- Electrochemistry (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Peptides Or Proteins (AREA)
Abstract
The present invention relates to a kind of detection method of contained polypeptide classes in pearl powder, it is heated to boiling and keeping the temperature and to obtain hydrolyzate under conditions of being stirred continuously the following steps are included: pearl powder is added to the water by (a)；(b) solid ammonium sulfate is added into the hydrolyzate, is stirred salting-out precipitate of saltouing to obtain；(c) salting-out precipitate is dissolved in NaCl solution, is then added in chromatographic column, carries out eluting to obtain eluent using the NaCl solution；(d) the characteristic peak quantity that detection obtains polypeptide is carried out to the eluent using ultraviolet specrophotometer；(e) separation is extracted to the eluent as solvent using n-Butanol acetic acid, is developed the color using ninhydrin and determines amount of speckle；When the amount of speckle is consistent with the characteristic peak quantity.So as to prove and explain pearl anti-aging pharmacological action.
Description
Technical field
The invention belongs to pearl powder fields, are related to a kind of detection method, and in particular to contained polypeptide kind in a kind of pearl powder
The detection method of class.
Background technique
Pearl is a kind of traditional rare Chinese medicine, has the history of two thousand years in China, have beauty treatment, beauty, anti-aging,
The various aspects effect such as tranquilizing the mind, myogenic.Composition contained by pearl is comprehensive more, and inhomogeneity composition role is also different, and pharmacological research proves
Polypeptide has anti-aging effects, it is therefore desirable to the quantity of polypeptide composition in pearl is determined and be separated, to verify pearl
Anti-aging effects.
Summary of the invention
A kind of inspection of contained polypeptide classes in pearl powder is provided the invention aims to overcome the deficiencies in the prior art
Survey method.
In order to achieve the above objectives, the technical scheme adopted by the invention is as follows: the inspection of contained polypeptide classes in a kind of pearl powder
Survey method, it the following steps are included:
(a) pearl powder is added to the water, is heated to boiling and keeping the temperature and to obtain hydrolyzate under conditions of being stirred continuously；
(b) solid ammonium sulfate is added into the hydrolyzate, is stirred salting-out precipitate of saltouing to obtain；
(c) salting-out precipitate is dissolved in NaCl solution, is then added in chromatographic column, is washed using the NaCl solution
Take off to obtain eluent；
(d) the characteristic peak quantity that detection obtains polypeptide is carried out to the eluent using ultraviolet specrophotometer；
(e) separation is extracted to the eluent as solvent using n-Butanol acetic acid, is developed the color using ninhydrin true
Determine amount of speckle；When the amount of speckle is consistent with the characteristic peak quantity.
Optimally, it further include:
(f) using the acetum of 0.1M as buffer to the eluent carry out electrophoresis detection, using ninhydrin develop the color with
Amount of speckle is checked again.
Optimally, in step (e), the quantity of the spot is two, RfAxis values are respectively 0.35,0.43.
Optimally, in step (a), the mass ratio of the pearl powder and the hot water is 1:1~3, and the soaking time is 1
~3h.
Optimally, in step (b), the ratio of the solid ammonium sulfate and the water is 10~20g:100ml.
Optimally, in step (c), the concentration of the NaCl solution is 0.5~1.0g/ml.
Due to the above technical solutions, the present invention has the following advantages over the prior art: in pearl powder of the present invention
The detection method of contained polypeptide classes by the chemical treatment such as being hydrolyzed, saltouing to pearl powder, and combines expansion separation, purple
The detection of outer spectrophotometer realizes appraising and deciding and separating to polypeptide classes, so as to prove and explain the anti-aging of pearl
Pharmacological action.
Detailed description of the invention
Fig. 1 is the map schematic diagram that UV spectrophotometer measuring obtains in the present invention.
Specific embodiment
The detection method of contained polypeptide classes in pearl powder of the present invention, it is the following steps are included: water is added in pearl powder by (a)
In, it is heated to boiling and keeping the temperature and to obtain hydrolyzate under conditions of being stirred continuously；(b) solid ammonium sulfate is added into the hydrolyzate,
It is stirred salting-out precipitate of saltouing to obtain；(c) salting-out precipitate is dissolved in NaCl solution, is then added in chromatographic column, using described
NaCl solution carries out eluting to obtain eluent；(d) detection is carried out to the eluent using ultraviolet specrophotometer and obtains polypeptide
Characteristic peak quantity；(e) separation is extracted to the eluent as solvent using n-Butanol acetic acid, it is aobvious using ninhydrin
Color determines amount of speckle；When the amount of speckle is consistent with the characteristic peak quantity.By the way that pearl powder is hydrolyzed,
It the chemical treatment such as saltouts, and expansion separation, the detection of ultraviolet specrophotometer is combined to realize appraising and deciding and divide to polypeptide classes
From, so as to prove and explain pearl anti-aging pharmacological action.
Above-mentioned detection method is further comprising the steps of: (f) using the acetum of 0.1M as buffer to the eluent into
Row electrophoresis detection is developed the color to check amount of speckle again using ninhydrin, can further determine that the type of polypeptide in this way, to protect
Demonstrate,prove the order of accuarcy of detection.In step (e), the quantity of the spot is two, RfAxis values are respectively 0.35,0.43.Step
Suddenly in (a), the mass ratio of the pearl powder and the hot water is 1:1~3, and the soaking time is 1~3h.In step (b), institute
The ratio for stating solid ammonium sulfate and the water is 10~20g:100ml.In step (c), the concentration of the NaCl solution is 0.5~
1.0g/ml。
Below in conjunction with embodiment, invention is further explained.
Embodiment 1
The present embodiment provides a kind of detection method of contained polypeptide classes in pearl powder, it the following steps are included:
(a) pearl powder is added to the water (mass ratio of pearl powder and water is 1:2), is heated under conditions of being stirred continuously
It boils (about 100 DEG C) and keeps the temperature 2h and obtain hydrolyzate；
(b) solid ammonium sulfate (ratio of ammonium sulfate and water is 15g:100ml) is added into hydrolyzate, is stirred and saltouts
Obtain salting-out precipitate；
(c) by salting-out precipitate be dissolved in NaCl solution (concentration of NaCl solution be 0.9g/ml, the ratio of salting-out precipitate and NaCl solution
Example is about 20g:100ml) in, it is then added in chromatographic column, carries out eluting to obtain eluent using the NaCl solution of same concentrations；
(d) the characteristic peak quantity (as shown in Figure 1) that detection obtains polypeptide is carried out to eluent using ultraviolet specrophotometer；
(e) it uses n-Butanol acetic acid (volume ratio 1:1) to extract separation to eluent as solvent, uses indenes
Triketone, which develops the color, determines that (amount of speckle is also two to amount of speckle, RfAxis values are respectively 0.35,0.43)；When amount of speckle with
Characteristic peak quantity (when amount of speckle and characteristic peak quantity are inconsistent, can also continue to following steps) when consistent；
(f) using the acetum of 0.1M as buffer to the eluent carry out electrophoresis detection, using ninhydrin develop the color with
Check amount of speckle again (the final type for determining polypeptide in pearl powder is two kinds).
Embodiment 2
The present embodiment provides a kind of detection method of contained polypeptide classes in pearl powder, in it and embodiment 1 basic one
It causes, unlike: in step (a), the mass ratio of pearl powder and water is 1:1, and heat preservation 1h obtains hydrolyzate；In step (b), to hydrolysis
Solid ammonium sulfate is added in liquid, so that the ratio of ammonium sulfate and water is 10g:100ml；In step (c), the concentration of NaCl solution is
0.5g/ml。
Embodiment 3
The present embodiment provides a kind of detection method of contained polypeptide classes in pearl powder, in it and embodiment 1 basic one
It causes, unlike: in step (a), the mass ratio of pearl powder and water is 1:3, and heat preservation 3h obtains hydrolyzate；In step (b), to hydrolysis
Solid ammonium sulfate is added in liquid, so that the ratio of ammonium sulfate and water is 20g:100ml；In step (c), the concentration of NaCl solution is
1g/ml。
The above embodiments merely illustrate the technical concept and features of the present invention, and its object is to allow person skilled in the art
Scholar cans understand the content of the present invention and implement it accordingly, and it is not intended to limit the scope of the present invention.It is all according to the present invention
Equivalent change or modification made by Spirit Essence, should be covered by the protection scope of the present invention.
Claims (6)
1. the detection method of contained polypeptide classes in a kind of pearl powder, which is characterized in that it the following steps are included:
(a) pearl powder is added to the water, is heated to boiling and keeping the temperature and to obtain hydrolyzate under conditions of being stirred continuously；
(b) solid ammonium sulfate is added into the hydrolyzate, is stirred salting-out precipitate of saltouing to obtain；
(c) salting-out precipitate is dissolved in NaCl solution, is then added in chromatographic column, elute using the NaCl solution
Eluent；
(d) the characteristic peak quantity that detection obtains polypeptide is carried out to the eluent using ultraviolet specrophotometer；
(e) separation is extracted to the eluent as solvent using n-Butanol acetic acid, is developed the color using ninhydrin and determines spot
Point quantity；When the amount of speckle is consistent with the characteristic peak quantity.
2. according to claim 1 in pearl powder contained polypeptide classes detection method, which is characterized in that it further include:
(f) electrophoresis detection is carried out to the eluent using the acetum of 0.1M as buffer, is developed the color using ninhydrin with again
Check amount of speckle.
3. according to claim 1 in pearl powder contained polypeptide classes detection method, which is characterized in that in step (e), institute
The quantity for stating spot is two, RfAxis values are respectively 0.35,0.43.
4. according to claim 1 in pearl powder contained polypeptide classes detection method, it is characterised in that: in step (a), institute
The mass ratio for stating pearl powder and the hot water is 1:1 ~ 3, and the soaking time is 1 ~ 3h.
5. according to claim 1 in pearl powder contained polypeptide classes detection method, it is characterised in that: in step (b), institute
The ratio for stating solid ammonium sulfate and the water is 10 ~ 20g:100ml.
6. according to claim 1 in pearl powder contained polypeptide classes detection method, it is characterised in that: in step (c), institute
The concentration for stating NaCl solution is 0.5 ~ 1.0g/ml.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201811086157.5A CN109283025A (en) | 2018-09-18 | 2018-09-18 | The detection method of contained polypeptide classes in a kind of pearl powder |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811086157.5A CN109283025A (en) | 2018-09-18 | 2018-09-18 | The detection method of contained polypeptide classes in a kind of pearl powder |
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| CN109283025A true CN109283025A (en) | 2019-01-29 |
Family
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| CN201811086157.5A Pending CN109283025A (en) | 2018-09-18 | 2018-09-18 | The detection method of contained polypeptide classes in a kind of pearl powder |
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| CN103923166A (en) * | 2014-05-05 | 2014-07-16 | 西华大学 | Separation and purification method of bamboo leaf antioxidative peptide |
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